Objective To establish a satisfactory lung metastasis model of human choriocarcinoma using severe combined immunedeficient(SCID)mice and explore the appropriate cell concentration for the model.Methods Forty SCID mice aged between 5-6 weeks were randomly difided into four groups.1×107 cells/ml ×0.1 ml.5×106 cells/ml×0.2 ml and 1×106 cells/ml×0.1 ml of human choriocarcinoma cells JEG-3 were respectively injected in SCID mice of experimental groups by lateral tail vein,the remain group was assigned to the control group.The status and weisht of mice were observed every three days.When these mice were being dying.the size and the number of the lesions of lung metastasis in every mouse were inspected with Micro CT.After Micro CT inspection,the SCID mice were executed dissected to note whether there were tumors on all organ surfaces witll naked eyes.then made pathological sections from the metastaticfoci of fresh lung tissues,and cultured primarily cells and purified cells and passaged cells isolated from the same metastastic foci.The pathological sections were observed under the microscope.The special antigen human chorionic gonadotropin-beta subunit(β-hCG)of the choriocareinoma cells was immunohistochemically detected in the pathological sections and the cells out of cultured primarily cells.The chromosomes of the cells out of cultured primarily cells were analysed.Results Of the group inoeutated 1×107 cells/ml×0.1 ml.all mice died when inoculating.In the group of 5×106 cells/ml×0.2 ml,when inoculating, 3 mice died; the remain 7 mice were being dying on ( 18. 0 ±2. 0) days after injection. 5 of them, there were 1 - 3 lesions of lung metastasis after Micro CT inspection in each mice, and the diameter of the tumors lesions reached 1.5 - 3.5 ram, which was choriocarcinoma confirmed by pathological sections.The special antigen β-hCG was detected by immunohistoehemical method in the pathological sections of pulmonary tissue with tumor and in the cells, which were purified and passaged from being cultured primarily cells isolated from metastastic foci of fresh lung tissues from the SCID mice. The chromosome numbers of these cells out of cultured primarily ceils were variety from 19 to 128, and medal numbers were variety from 70 to 79. Conclusions We successfully established the lung metastatic model of human choriocarcinoma in SCID mice by injecting JEG-3 cells into lateral tail vein, of which 5 × 106 cells/ml × 0. 2 ml is the suitable concentration and volume for the model.

Objective To establish the high-performance liquid chromatography(HPLC)method for measuring concentrations of methotrexate(MTX)in rat lung and some other tissues through internal iliac artery infusion.Methods Fifly female Sprague-Dawley rats were included in this study.The rats were randomly assigned to two groups.Methotrexate was injected to group one through internal iliac artery,and was injected to group two through femoral vein.Blood and tissues were collected in each group at 15,30,60.90 and 120 minutes for detection of the drug concentrations with HPLC.Results The area under the concentration time curve(AUC)in rat lung,ovary and uterus in the artery group were separately(3.77±0.28),(4.40±0.40),(9.97±0.89)μg·h-1·g-1,which were significantly different from those of the vein group[(2.31±0.25),(3.91±0.19),(7.65±1.54)μg·h-1·g-1;P<0.05].The AUC in the rat plasma,heart,kidney,liver and spleen in the artery group were separately(6.13±0.53),(1.90 ±0.11),(5.32±0.89),(14.16±1.96),(0.76±0.20)μg·h-1·g-1.There were no significant differences from the vein group[(5.79±0.71),(1.64±0.29),(5.15±1.69),(14.29 ±3.47),(0.76±0.13)μg·h-1·g-1;P>0.05].Conclusions Through internal iliac artery infusion,there are higher drug concentrations in lung.uterus and ovarian compared to venous injection.The internal-arterial chemotherapy may be used to treat pulmonary metastasis of gynecological tumor.

Objective To investigate the NAD+ - dependent protein deacetylase SIRT1 expression in the cochlea of C57BL/6 mice ,a mouse model of age - related hearing loss(AHL) .Methods A total of 46 C57BL/6 mice were used ,and were divided into 2 groups ;according the age ,there were young group (1 ~ 2 months old ,23 mice) and old group (12 ~ 16 months old ,23 mice) .ABR measurements were performed on young and old C57BL/6 mice . The expression of SIRT1 in the cochlea was detected by qRT - PCR and immunofluorescence .Results The ABR thresholds in the old mice(4 kHz :82 .7 ± 7 .32 dB SPL ,8 kHz :80 .9 ± 7 .8 dB SPL ,16 kHz :89 .3 ± 5 .5 dB SPL ,32 kHz :89 .9 ± 4 dB SPL) were significantly higher than those in the young C57BL/6 mice(4 kHz :52 .1 ± 8 .3 dB SPL ,8 kHz :40 .5 ± 6 .1 dB SPL ,16 kHz :50 .7 ± 9 .4 dB SPL ,32 kHz :57 .6 ± 11 .9 dB SPL)(P < 0 .001) .SIRT1 mRNA expression was significantly decreased in the cochlea of old C57BL/6 mice in comparison with young mice ( P <0 .01) .SIRT1 protein was abundantly expressed in the inner hair cells ,outer hair cells ,supporting cells ,strial marginal cells ,strial intermediated cells ,and spiral ganglion neurons .The positive area and the average flourescence intensity of SIRT1 protein were reduced in old C57BL/6 mice(P< 0 .001) .Conclusion The down - regulation of SIRT1 mRNA and protein expression in the older C57BL/6 mouse cochlea may be correlated with the pathogenesis of AHL .

Objective To evaluate the value of spiral CT and 3D reconstruction in diagnosis and treatment of tibial plateau fractures.Methods 25 cases with tibial plateau fractures were examined with plain radiograhs,spiral CT and 3D reconstruction during July,1998 to December,2000.The tibial plateau fractures were classified according to Schatzker′s classification on the X-rays and 3D reconstruction images.On the superior view the tibial plateau fractures were divided into anterior lateral,posterior lateral,anterior medial and posterior medial fractures.Results 3D reconstruction images of 25 plateau fractures were the same as what arthroendoscopy were.25 fractures(including 23 arthroscopic management fractures)were operated under 3D reconstruction guide.No one case was infected.The treatment was valid.Conclusion Spiral CT and 3D reconstruction can offer more accurate classification and complement to new spatial classification of fractures.Also can provide better planning and management for the tibial plateau fractures.

Objective To evaluate the treatment of undercorrection after the excimer laser in situ keratomileusis (LASIK) for correcting moderate and high myopia. Methods An enhancement ablation was performed in 48 eyes of 39 patients who had undergone LASIK but remained in undercorrection. Four procedures were performed within 1 month postopoeratively, and the others performed between 3 and 10 months. The surgical technique includes the re-invert of the corneal cap from the temporal side, the excimer laser ablation, and the re-position of the cap.Results The undercorrection (spherical equivalent) ranged from -2.00 to -11.00 D, with a mean of -4.34D±1.95 D. Following up after enhancement ablation was done after 4 to 12 months, the refractions in the 42 eyes were found to be within ±1.00 D. Undercorrection of -2. 50 D to -5.00 D recurred in 6 eyes. Uncorrected visual acuity equals to the preoperative spectacle corrected visual acuity in 39 of 48 eyes (81.3%). Five eyes gained 1 line, 1 eye gained 2 lines and 4 eyes lost 1 line. No eyes had haze.Conclusion Undercorrection after LASIK can be corrected by an enhancement ablation of the stroma under the primary corneal cap with a 193 nm ArF excimer laser, and the time for the enhancement of ablation is at 3 months postoperatively.

Objective To measure the expression of activin receptor-like kinases 1(ALK1)in dermal fibroblasts from patients with systemic scleroderma(SSc)and to estimate its role in the production of fibronectin and plasminogen activator inhibitor-1(PAI-1).Methods Dermal fibroblasts were isolated from the lesions of 12 patients with SSc as well as the normal skin of 14 healthy controls,and subjected to a primary culture.The third-passage fibroblasts were used in the next experiment.Western blot and indirect immunofluorescence technique were utilized to quantify the expression of ALK1.A specific siRNA targeting ALK1 was designed,constructed,and transiently transfected into the control dermal fibroblasts,which were then classified into 2 groups to be cultured with or without the presence of transforming growth factor(TGF)-β1 for 72 hours followed by the detection of fibronectin and PAI-1 expression with Western blot.Results As Western blot and direct immunofluorescence technique showed,both control and SSc fibroblasts showed an expression of ALK1 in the cytoplasm and membrane,and the expression intensity of ALK1 in SSc fibroblasts was significantly higher than that in the control fibroblasts(1.97 ± 0.05 vs.1.12 ± 0.03,t =50.96,P ＜ 0.05).The expression of ALK1,fibronectin and PAI-1 was decreased by 90％,58％ and 31％ respectively in specific siRNA-transfected SSc fibroblasts compared with the control siRNA-transfected fibroblasts.TGFβ1 significantly increased the expression of ALK1,fibronectin and PAI-1 in the control siRNA-transfected fibroblasts,but the increase was markedly inhibited by the siRNA-targeting ALK1.Conlusion TGFβ1 can promote the production of fibronectin and PAI-1 via ALK1 in fibroblasts,and ALK1 may be involved in the development of sclerosis in SSc.

Objective To probe the relationship between the patients′ prognosis and the gene of multidrugs and drug resistance in uveal melanoma. Methods The gene expression of cyclin D1, epithelial growth factor receptor (EGFR), non metastasis gene 23 (nm 23), P glucose protein (P gp), multidrug resistance relation protein (MRP) and lung resistance protein (LRP) expression in 96 cases of uveal melanoma were detected by depigment immunohistochemistry. The patients with complete anamnesis data were observed continuously, and the follow up results were classified. Results Among the 96 cases of uveal melanoma, the epithelioid cell type was in 21, the mixed cell type in 56, and the spindle cell type in 19; including 76 at intraocular stage and 20 at extraocular stage. As the level of metastasis suppress gene nm 23 expression decreased and the level of cyclin D1 and EGFR expression increased, the expression level of drug resistance genes increased. The levels of LRP and MRP had negative correlation to the expressions of nm 23 and postive correlation to the expressions of nm 23, Cyclin D1 and EGFR. In 58 patients′ who were observed continuously, 19 died in 5 years and 26 survived over 5 years. Conclusion There are significant association between patients′ prognosis and multidrug and drug resistance gene in uveal melanoma.

Objective: To analyze the relevant literature regarding the effect of tuina on postpartum milk secretion and thus summarize the clinical rules on tuina for lactation disorder. Method: Investigate the relationship between tuina and postpartum milk secretion for four aspects, including the initial time of lactation, level of serum prolactin, volume of lactation, and Chinese medicine's understanding of tuina on milk secretion. Result and Conclusion: Tuina on breasts after childbirth can speed and promote lactation. This has been proved by clinical practice over the past hundreds of years, along with modern laboratory and scientific research. This method, therefore, is of great significance in obstetrical nursing.

Objective To investigate vitamin D ( VD) level and its influencing factors in 4-15-year-old children in southwest of Zhejiang province, so as to guide intervention strategy for improving VD level. Methods A total of 2 620 4 -15-year-old healthy children who visited Lishui People's Hospital for regular physical check-up between January and December 2014 were divided into 4 age groups, namely 4 -6 years (kindergarten), 7 -9 years (lower grades in primary school), 10 -12 years (higher grades in primary school), and 13-15 years (junior middle school).We randomly selected 200 children from each age group using a random number table.A survey using self-developed questionnaire was conducted to obtain information regarding diet, exposure to sunlight, and VD supplement.Serum 25-( OH) D level was measured with electro-chemiluminescence.VD nutritional status and composition, diet, sunlight exposure, VD supplement were com-pared among the groups.Results In southwest Zhejiang, the mean VD level in primary and middle school students was ( 24.76 ±8.39 ) μg/L, clinical deficiency rate was 31.5%, subclinical deficiency rate was 45.0%, and only 23.5% reached the appropriate level, which was even lower ( 14.5%) in junior middle school students.The VD deficiency rate elevated significantly along with the growth of age (P=0.036).The level of VD was positively correlated with sunlight exposure and VD supplementation, and showed significant in-ter-group differences ( all P<0.05 ) .Conclusions The nutritional status of VD is unfavorable and sunlight exposure insufficient in primary and middle school students in southwest Zhejiang province.We should encour-age students to take more outdoor activities and increase sunlight exposure to improve their VD nutritional sta-tus.Children >7 years should be given more attention to in the intervention of increasing sunlight exposure. Students with difficulties in getting adequate sunlight exposure need VD supplements.

[Objective] To investigate the efficacy of Xiangsha Yangwei Wan (XYW) on the inhibition of chemotherapy-induced chemical gastritis and upper gastrointestinal symptoms. [Methods] A randomized controlled trial (RCT) was carried out in 87 cases of maligant tumor confirmed by histopathological and cytological examination. All the cases were treated with chemotherapy regimen of adriamycin ( ADM) , adcarbazine ( DTIC) , platinum-based agents, irinotecan (CPT-11) and fluorouraeil (5-FU). Meanwhile, 41 cases in group A were also treated with dexamethason and 5-HT3 receptor antagonist as an essential anti-emesis and with XYW to regulate gastrointestinal function; other 46 cases in group B were given dexamethason and 5-HT3 receptor antagonist only. Seven days after treatment, incidences of chemical gastritis and upper gastrointestinal symptoms were observed and compared between the two groups. [Results] Seven days after chemotherapy, upper gastrointestinal symptoms and signs were relieved in 95.1% (39/41) of the patients of group A and in 76.1 % (35/46) of group B; the occurring rate of chemical gastritis with symptoms was 2.4% (1/41) in group A and 19.6% (9/46) in group B, the differences being significant statistically between the two groups( P - 0.0122) . [ Conclusion ] XYW has an inhibitory effect on chemotherapy-induced chemical gastritis and upper gastrointestinal symptoms.